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Lecture 4

MCB 2000 Lecture Notes - Gonorrhea, Pelvic Inflammatory Disease, Chromosome
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3 Pages
39 Views
Summer 2018

Department
Microbiology and Cell Science
Course Code
MCB 2000
Professor
Abdolkarim Asghari
Lecture
4

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Lecture 4
Microscopy
o Magnification ranges
In order from strongest magnification to weakest
Atomic force microscope
Transmission electron microscope
o Can show details of the inside of the microbe
Scanning electron microscope
o Can only show details of the surface of the microbe
Light microscope
o Have to use electron microscope to see viruses
o Brightfield = use visible light as source of illumination (light
colored background)
o Darkfield = illuminates background, uses special
condenser with opaque disk, ligh is reflected by specimen
(black background)
o Phase-contrast = shows more details than other light
microscopes
Unaided eye
o Other types of microscopes
Confocal = uses a single photon to illuminate one plane of a specimen at
a time
Will have a black background with many different colors
o Immunofluorescence = use antibodies made by the immune system to recognize
pathogens
Diagnostic tool
Tope of Y shape of antibodies binds to infection
Tails of antibodies are dyed and illuminate the pathogens they attach to
Antibodies are produced in the spleen
Monoclonal antibody = technique used to produce one type of antibody
Gram staining
o Process
Application of crystal violet (purple dye)
Gram positive bacteria stain purple because they have a thick cell
wall that this dye sticks to
Initially stains all bacteria
Application of iodine (mordant)
Alcohol wash (decolorization)
Washes out any bacteria that can’t hang (gram negative)
Application of safranin (counterstain)
Stains gram negative bacteria a red/pink color because they have
a thin cell wall and will not retain purple dye after alcohol wash
o Gram pos and neg bacteria have to be treated differently in healthcare so this is
very important for identification of the bacteria type
Medically important structural stains
o Capsule stain = stains the capsule around the bacteria and allows for ID of the
capsule
Bacteria that have an extra layer around them
Are harder to kill because they have more protection
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Description
Lecture 4 Microscopy o Magnification ranges In order from strongest magnification to weakest Atomic force microscope Transmission electron microscope o Can show details of the inside of the microbe Scanning electron microscope o Can only show details of the surface of the microbe Light microscope o Have to use electron microscope to see viruses o Brightfield = use visible light as source of illumination (light colored background) o Darkfield = illuminates background, uses special condenser with opaque disk, ligh is reflected by specimen (black background) o Phasecontrast = shows more details than other light microscopes Unaided eye o Other types of microscopes Confocal = uses a single photon to illuminate one plane of a specimen at a time Will have a black background with many different colors o Immunofluorescence = use antibodies made by the immune system to recognize pathogens Diagnostic tool Tope of Y shape of antibodies binds to infection Tails of antibodies are dyed and illuminate the pathogens they attach to Antibodies are produced in the spleen Monoclonal antibody = technique used to produce one type of antibody Gram staining o Process Application of crystal violet (purple dye) Gram positive bacteria stain purple because they have a thick cell wall that this dye sticks to Initially stains all bacteria Application of iodine (mordant) Alcohol wash (decolorization) Washes out any bacteria that cant hang (gram negative) Application of safranin (counterstain) Stains gram negative bacteria a redpink color because they have a thin cell wall and will not retain purple dye after alcohol wash o Gram pos and neg bacteria have to be treated differently in healthcare so this is very important for identification of the bacteria type Medically important structural stains o Capsule stain = stains the capsule around the bacteria and allows for ID of the capsule Bacteria that have an extra layer around them Are harder to kill because they have more protection
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